Journal: bioRxiv
Article Title: Fission yeast histone chaperone Rtt106 regulates histone levels, prevents early division, and promotes genome stability
doi: 10.1101/2025.11.01.686071
Figure Lengend Snippet: A) Strains were assessed for drug sensitivities using a spot test on solid YES medium. Strains were normalized at 5 x 10 6 cells/mL and diluted in four 1:5 dilutions. The rtt106 Δ grew similarly to wild type (wt) under all drug conditions (hydroxyurea, camptothecin, and phleomycin). Double mutant checkpoint strains with rtt106 Δ had similar drug sensitivities to their single mutant checkpoint parental strain. B,C) Growth in YES liquid cultures was modeled by monitoring OD600 at a range of hydroxyurea (B) and camptothecin (C) doses. Shown are dose-response curves for each strain. A minimum of 3 experimental replicates were normalized to respective no drug value, and then modeled with a log(inhibitor) response and variable slope. Shown is the composite response across replicate data for each strain and condition. In HU, matched rtt106 Δ strains behaved like the corresponding rtt106+ strain. However, variation in rad3 Δ rtt106 Δ versus rad3 Δ indicated that subtle changes occur in HU that are not captured by OD600 liquid cultures. C) IC50 curves in CPT are similar in wild type, cds1 Δ, and chk1 Δ pairs ( rtt106 + vs rtt106 Δ ). The rad3 Δ rtt106 Δ cells grow better in CPT than rad3 Δ alone (p=0.016). IC50 values are presented in Table XX. D) Area under the curve (AUC) was used to model differences in replicate growth curves. HU decreases growth in all matched rtt106+ vs rtt106 Δ pairs. CPT effects on growth curves are not significant in wild type, cds1 Δ, and chk1 Δ. An exception is that rad3 Δ rtt106 + cells are more sensitive to CPT than rad3 Δ rtt106 Δ, which has a significant impact in AUC (** p<0.01).
Article Snippet: Drug plates were made freshly in YES agar medium using filter sterilized drug stocks including: hydroxyurea (1M in water, BioShop, Canada); camptothecin (10 mM in DMSO; BioShop Canada); phleomycin (100 mg/mL in PBS, Invitrogen Zeocin).
Techniques: Spot Test, Mutagenesis
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